Leman SENCAR, Derviş Mansuri YILMAZ, Dilek GÖKTÜRK, Sema ÖZANDAÇ POLAT, Gülfidan COŞKUN, Dilek ŞAKER, Tuğçe SAPMAZ, Samet KARA, Alper ÇELENK, Sait POLAT

Glioblastoma hücre hattında (U87) siklopamin ve temozolomid kombine tedavisinin miR-20a ekspresyonu üzerine etkileri

Amaç: Bu çalışmada, glioblastoma hücre hattında (U87) siklopamin ve temozolomid (TMZ) tedavisinin ayrı ayrı ve kombine olarak uygulanmasından sonra miR-20a ekspresyonunun araştırılması ve bu miRNA’nın glioblastoma tedavisindeki etkinliğinin değerlendirilmesi amaçlanmıştır. Gereç ve Yöntem: U87 MG, tedaviden önce 35 mm'lik kültür kabı üzerine yerleştirildi ve çoğaltıldı. Çalışma kapsamında 7 grup oluşturuldu: Grup 1: Kontrol grubu, Grup 2: Sham grubu (Dimetil sülfoksit), Grup 3: 2 Gün TMZ, Grup 4: 5 Gün TMZ, Grup 5: 3 saat siklopamin, Grup 6: 2 Gün TMZ + 3 saat siklopamin, Grup 7: 5 Gün TMZ + 3 saat siklopamin. Tedaviden sonra total miRNA izolasyonu ve qRT-PCR yapıldı. Bulgular: Glioblastoma hücre hattında miR-20a ekspresyonunun grup 3, grup 4, grup 5, grup 6 ve grup 7’de belirgin olarak azaldığı tespit edildi. Ancak bu azalmanın en fazla grup 7 de olduğu saptandı . Ek olarak grup 7’de hücre sayısının azaldığı görüldü. Sonuç: Glioblastoma hücrelerinde miR-20a’nın yüksek oranda eksprese olduğu; ancak siklopamin ve TMZ tedavilerinden sonra ekspresyonun azaldığı tespit edildi. miR-20a’nın glioblastoma için yeni bir terapötik hedef ve biyobelirteç olarak kullanılabileceği, siklopamin ve TMZ’nin glioblastomada tedavi amaçlı kullanılabileceği ancak bu konuda daha ileri çalışmaların yapılması gerektiği sonucuna varıldı.

Anahtar Kelimeler:

Glioblastoma, siklopamin, temozolomid, MiR-20a.

Effects of cyclopamine and temozolomide combined treatment on miR-20a expression in glioblastoma cell line (U87)

Purpose: The aim of this study was to investigate miR-20a expression in glioblastoma cell line (U87) after applying cyclopamine and temozolomide (TMZ) treatment separately and in combination and to evaluate the efficiency of miR-20a in glioblastoma treatment. Materials and Methods: U87 MG was placed on a 35 mm culture dish before treatment. Within the scope of the study, 7 groups were formed: Group 1: Control group, Group 2: Sham group (Dimethyl sulfoxide), Group 3: 2 Days TMZ, Group 4: 5 Days TMZ, Group 5: 3 Hours Cyclopamine, Group 6: 2 Days TMZ + 3 Hours Cyclopamine, Group 7: 5 Days TMZ + 3 Hours Cyclopamine Total miRNA isolation and qRT-PCR was performed after treatment. Results: MiR-20a expression in the glioblastoma cell line was found to be significantly decreased in group 3, group 4, group 5, group 6 and group 7. However, it was determined that this decrease was highest in group 7. In addition, it was observed that the number of cells decreased in group 7. Conclusion: MiR-20a is highly expressed in glioblastoma cells; however, it was determined that the expressions decreased after Cyclopamine and TMZ treatments. miR-20a can be used as a new therapeutic target and biomarker for glioblastoma, Cyclopamine and TMZ can be used for treatment in glioblastoma, but further studies are needed on this subject.

Keywords:

Glioblastoma, cyclopamine, temozolomide, MiR-20a.,

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